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Phenotypic and functional characterization of two bovine mammary epithelial cell lines in 2D and 3D models.

Authors
  • Arévalo Turrubiarte, Magdalena1
  • Perruchot, Marie-Hélène1
  • Finot, Laurence1
  • Mayeur, Frédérique1
  • Dessauge, Frédéric2
  • 1 Institut National de la Recherche Agronomique, UMR1348 Pegase, Saint-Gilles, France; and Agrocampus Ouest, UMR1348 Pegase, Rennes, France. , (France)
  • 2 Institut National de la Recherche Agronomique, UMR1348 Pegase, Saint-Gilles, France; and Agrocampus Ouest, UMR1348 Pegase, Rennes, France [email protected] , (France)
Type
Published Article
Journal
AJP Cell Physiology
Publisher
American Physiological Society
Publication Date
Mar 01, 2016
Volume
310
Issue
5
Identifiers
DOI: 10.1152/ajpcell.00261.2015
PMID: 26659725
Source
Medline
Keywords
License
Unknown

Abstract

Immortalized bovine mammary epithelial cells (BME-UV1) and immortalized bovine mammary alveolar cells (MAC-T) have been extensively used as in vitro cell models to understand milk production in dairy cows. Precise knowledge about their phenotype and performance remains, however, unknown. This study aims to characterize MAC-T and BME-UV1 profiles when cultured in two-dimensional adherent, three-dimensional adherent (Matrigel), and three-dimensional no adherent [ultralow attachment (ULA)] supports. MAC-T and BME-UV1 were compared according to their proliferation capacities and to specific cell surface markers CD24, CD326 [epithelial cell adhesion molecule (EpCAM)], CD10, and integrin CD49f (α-6). Cytokeratin (CK14 and CK19), signal transducer and activator of transcription 5, and other proteins (occludin and cadherin-1) were analyzed. BME-UV1 in ULA support expressed higher CD49f marker. A different intensity of CD49 staining allowed the discrimination between the two cell lines in adherent condition. CD10, EpCAM, and CK19 expressions show that BME-UV1 cells have luminal capacity, while MAC-T has a myoepithelial profile with a high expression of CK14. BME-UV1 cells possess a closer committed progenitor profile due to their higher expression in aldehyde dehydrogenase and EpCAM. We observed that BME-UV1 cells have a better capacity to form spherical structures, mammospheres, in Matrigel than MAC-T, which was confirmed by the higher mammosphere area. In the ULA condition, BME-UV1 proliferated over the 6 days of culture. Taken together, our results clearly confirm the BME-UV1 luminal profile and MAC-T ductal/myoepithelial-like phenotype.

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